System and method for remote navigation of a specimen

ABSTRACT

A method of providing remote navigation of a specimen includes receiving a command to remotely control a microscope and a camera coupled with the microscope to view a portion of a specimen disposed on the microscope, capturing a digital image of the portion with the camera in response to the command, representing the digital image as a plurality of digital image components, each of the digital image components providing a different level of detail of the image, and transmitting the components in sequential order of increasing level of detail from least detailed to most detailed.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.12/118,356, filed May 9, 2008, which is a continuation of U.S. patentapplication Ser. No. 11/485,478, filed Jul. 12, 2006, which is adivisional of U.S. patent application Ser. No. 10/448,913, filed May 30,2003, now U.S. Pat. No. 7,224,839, which is a continuation of U.S.patent application Ser. No. 09/323,371, filed Jun. 1, 1999, now U.S.Pat. No. 6,606,413, which claims benefit of U.S. Provisional ApplicationNo. 60,087,523, filed Jun. 1, 1998, each of which are hereby fullyincorporated herein by reference.

FIELD

The present invention relates generally to the field of remote operationand viewing of a videographic imaging system and, more particularly, tothe acquisition and transmission of images useful in the field oftelemedicine to effect remote site diagnostic and consultationprocedures.

BACKGROUND

The digital revolution has intruded in almost every sphere of modernelectronic communications and has given rise to applications andabilities that were not even considered prior to the introduction of thesmall platform computer system in the late 1960's and its subsequentdevelopment through the 70's, 80's and 90's. Although totally pervasivein every aspect of society and sector of the economy, the digitalrevolution has had a significant impact in the field of electroniccommunication and, most particularly, to that area relating to thecapture, transmission and faithful reproduction of audiographic andvideographic data. No one field has benefited more from the capabilitiesgenerated by the digital revolution than that of telemedicine.

Functionally, telemedicine allows a physician to have a remote sitecapability by means of which they are able to direct procedures, makediagnosis, and generally engage in the practice of certain forms ofmedicine without the need to be physically present in the operatingtheater or the examination room in order to effect a practicallyreal-time interaction. In the field of pathology, specifically,telemedicine (telepathology to be more precise) allows the investigatingpathologist to be separated from the local origin of tissue to beinvestigated and, ideally, still be able to make an effectiveinvestigation of a tissue sample in order to render a diagnosticopinion.

The current trend in telemedicine in general, and telepathology inparticular, gives rise to some very interesting implications for thequality of healthcare services available to the public at large.Telepathology particularly allows a surgeon about to perform an invasivesurgical procedure, to select a particular specialist without regard tothat specialist's location. Selection of pathology services need only bemade, therefore, by determining those most suited, or experienced, indealing with the particular organ system under consideration. In thedaily routine of a large hospital, such service flexibility becomeshighly relevant when the diverse character of the procedures carried outas such hospitals is considered. Likewise, small to medium sizedhospitals, which may not be able to support a large pathology staffincorporating the many subspecialties required for full coverage, areable to avail themselves of the same quality of pathology services thatone might find in a major urban hospital. Clearly, the benefits oftelemedicine, particularly telepathology, would be most greatly felt bysmall to medium sized hospitals in remote areas of the country where thesize and quality of specialty medical staff is necessarily limited todue to geographical isolation.

The enabling tool for providing telepathology services is atelemicroscopy system connected to a bi-directional telecommunicationsnetwork which is pervasive enough to allow the necessary equipment to beset up and operated virtually anywhere. Conventional forms oftelemicroscopy equipment are generally well known in the art andsuitably comprise a remote controlled microscope system where microscopeimages are acquired with a conventional video camera and transmitted,for display, to a control system. Remote operation of the microscopesystem and remote display of transmitted images can be realisticallyperformed using a variety of communications technologies. However, inorder to ensure a general availability of a developing telepathologynetwork, interconnectivity is most realistic in the context of narrowband or broadband landline connections. Narrow band systems (like PSTNand ISDN) generally guarantee worldwide availability for very low costs,but at the price of bandwidth and/or channel capacity. Broadband systems(like ATM) allow enhanced channel capacity but still suffer from a lackof sufficient bandwidth to allow video transmissions at anythingapproximating real-time. Because of these limitations, conventionaltelemicroscopy systems have had to make certain compromises betweenchannel capacity and image quality. The higher the quality of thetransmitted image, the longer the time it takes to complete atransmission. Conversely, when transmission speed is an overridingconcern, image quality necessarily suffers.

With a limited bandwidth available on PSTN (Public Switched TelephoneNetwork) and ISDN (Integrated Services Digital Network), the only meansavailable to increase transmission speed is to reduce the average numberof transmitted bits per image, i.e., compress the digital image datadeveloped by video camera. Before telepathology services become trulyviable, image transmission must operate at bit rates of only a fewhundred kilobits or a few megabits per second, which can only beachieved through rather large compression of the data.

Most sensory signals contain a substantial amount of redundant orsuperfluous information. For example, a conventional video camera, thatcaptures approximately 30 frames per second from a stationary image,produces very similar frames, one after the other. Compressiontechniques attempt to remove the superfluous information from repetitiveframes, such that a single frame can be represented by a reduced amountof finite data, or in the case of time varying images, by a lower datarate. It is well known in the art that digitized video signals comprisea significant amount of statistical redundancy, i.e., samples aresimilar to each other such that one sample can be predicted fairlyaccurately from another. By removing the predictable or similaritycomponent from a stream of samples, the video data rate can be reduced.Such statistical redundancy is able to be removed without perturbing theremaining information. That is, the original uncompressed data is ableto be recovered almost exactly by various inverse operations. Thealgorithms used in a compression system depend on the availablebandwidth, the features required by the application, and theaffordability of the hardware required for implementation of thecompression algorithm on both the encoding and decoding side.

BRIEF SUMMARY

In an aspect, a method of providing remote navigation of a specimenincludes receiving a command to remotely control a microscope and acamera coupled with the microscope to view a portion of a specimendisposed on the microscope, capturing a digital image of the portionwith the camera in response to the command, representing the digitalimage as a plurality of digital image components, each of the digitalimage components providing a different level of detail of the image, andtransmitting the components in sequential order of increasing level ofdetail from least detailed to most detailed.

In another aspect, a system providing remote navigation of a specimenincludes a processor that executes instructions and thereby causes theprocessor to: receive a command to remotely control a microscope and acamera coupled with the microscope to view a portion of a specimendisposed on the microscope, capture a digital image of the portion withthe camera in response to the command, represent the image as aplurality of digital image components, each of the digital imagecomponents providing a different level of detail of the image, andtransmit the components in sequential order of increasing level ofdetail from least detailed to most detailed.

In a further aspect, a method of navigating a slide includestransmitting at least one command to remotely view a portion of aspecimen, and receiving an increasingly more detailed image of theportion.

In yet another aspect, a system for navigating a slide includes aprocessor that executes instructions and thereby causes the processor totransmit at least one command to remotely view a portion of a specimen,and receive an increasingly more detailed image of the portion.

BRIEF DESCRIPTION OF THE DRAWINGS

The particular features, aspects and advantages of the present inventionwill be more fully understood when considered with respect to thefollowing detailed description and accompanying drawings wherein:

FIG. 1 is a semi-schematic black level diagram of a host network serverplatform, including a telemicroscopy system in accordance with practiceof principles of the present invention;

FIG. 2 is a semi-schematic block level diagram of a network clientsystem useful for hosting a remote site telepathology application inaccordance with practice of principles of the present invention;

FIG. 3 is an exemplary illustration of microscope sample stagetranslational motion defining previously transmitted and new portions ofa telemicroscope field of view;

FIG. 4 is a semi-schematic simplified block diagram of one embodiment ofa video image transmission system including multi-tiered compressionaccording to the present invention;

FIG. 5 is a semi-schematic simplified block diagram of a decompressionsystem adapted to receive compression packaged video image transmissionsin accordance with the invention;

FIG. 6 is a semi-schematic simplified block diagram of an additionalembodiment of a layered decompression system in accordance with theinvention; and

FIG. 7 is a semi-schematic simplified time verses image quality graphdepicting the relationship of image detail with viewer perception inaccordance with the invention.

DETAILED DESCRIPTION OF THE DRAWINGS

In order to gain a complete understanding of the compression packagedimage transmission system and method of the present invention, it willbe useful to examine how the system might function in the context of atypical telepathology procedure. During the course of a surgicalintervention, as a surgeon is preparing to perform an invasiveprocedure, the surgeon will typically remove a sample of tissue from apatient and forward the tissue sample to the hospital's diagnosticlaboratory for immediate evaluation. The tissue sample is prepared inconventional fashion and loaded onto the sample stage of an examiningmicroscope, comprising the laboratory's telemicroscopy system, where animage of the tissue sample is captured by a video camera andelectronically communicated to a pathologist at a remote site forevaluation. During the initial, or preliminary, evaluation, apathologist is able to view the tissue sample in macro and is furtherable to give directions to laboratory personnel as to location anddirection of sectioning to be performed in order to further anysubsequent diagnosis. Frozen sections are prepared according to theguidelines of the pathologist, the sections are mounted on glass slides,appropriately stained, and subsequently loaded onto a roboticallycontrolled microscope stage of the telemicroscopy system. Control of thetelemicroscopy system is then given to the remote-site pathologist.

The remote site pathologist is able to manipulate all of themicroscope's features and view images appearing under the microscopeobjective as though the pathologist were present at the laboratory siteand directly manipulating the microscope. Motion control of themicroscope stage in X-Y directions, as well as focus control of thestage in the Z direction, is performed by issuing the appropriatecontrol commands to a remote site small platform computer system which,in turn, transmits motion control commands to the telemicroscopy systemin the hospital laboratory. Following each X and/or Y translationalmovement of the microscope stage, the resulting tissue sample image istransmitted to the pathologist's remote site, where it is displayed on ahigh resolution monitor. The pathologist is, thus, able to manipulateand view a tissue sample as if the microscope containing the specimenwere directly in front of him.

Following the pathologist's investigation, the pathologist can prepare areport which provides the surgeon with his diagnostic opinion in thecase, either over the same bi-directional communication medium used toexamine the specimen, or by any one of a number of various otherelectronic communication forms available to the pathologist. Inparticular, the pathologist may transmit a written protocol to thesurgeon by facsimile, e-mail, and the like, or make a direct oral reportto the surgeon and follow-up with written documentation communicatedelectronically. After making a diagnosis, the pathologist releases thetelemicroscopy connection and is then available for a next consultationwith some other surgery team which might be preparing to perform adifferent surgical procedure in a totally different location. In themeantime, the original consulting surgeon is able to continue theintervention procedure according to the outcome of the diagnosis made bythe pathologist. As a check, the original consulting surgeon may decideto forward the original tissue specimen to a local pathology laboratoryfor a final diagnosis, in accordance with conventional acceptedprocedure. This manner of “Gold Standard” cross-checking, isparticularly useful as a means of acquiring data on the accuracy ofresults obtained by formulating diagnostic opinions on the basis oftelevised images which have been compressed, transmitted over longdistances over relatively “noisy” communication connections,decompressed and viewed on a high resolution video monitor screen.

In this regard, it bears mentioning that if visual image data can betransmitted to a remote site pathologist for the purpose of obtaining adiagnostic opinion, the same visual image data can be transmitted to aconsulting pathologist, by either the primary diagnostician, or thehospital. A primary and a consulting pathologist, or more than oneconsulting pathologist, are able to confer with respect to the samevisual image data representing the tissue sample. The ability to obtainon-line consultations is extremely advantageous, particularly where theproposed intervention implies a tissue morphology requiring the servicesof a pathologist or pathologists having a highly developed andcorrespondingly rare sub-specialty.

Turning now to FIG. 1, there is depicted a simplified semi-schematicblock diagram of an exemplary host or server telemicroscopy systemuseful in the practice of the present invention. FIG. 1 illustrates theprimary components of a remote, or robotically, controllabletelemicroscope, operable under software program control which would behosted on a control processor such a small platform personal computersystem. In accordance with the invention, telemicroscopy equipmentconnected to a telecommunication net in accord with a bi-directionalcommunication protocol, forms a key enabling tool for establishingeffective telemedicine services. This allows the system to combine thehigh-resolution and color saturation integrity of digital, still videoimages with the ability to establish bi-directional communicationbetween the “server” and a “client” system to enable remote, clientcontrol of the microscope in real time. Acquisition and transmission ofhigh-resolution video images of desired portions of a specimen can beperformed in a time period consistent with hands-on, real-time opticalpractice.

The telemicroscope portion, indicated generally at 10 suitably comprisesa remotely controllable microscope 12 configured with an illuminated,robotically controllable microscope stage 14. The microscope stage ismovable in X and Y directions and is controllable from a remote sourceby mechanically coupling X and Y translation motors to the stageplatform through control circuitry 16. A suitable illumination source isdisposed beneath the stage and is also translationally movable beneaththe stage in order to shift the apparent illumination source withrespect to a specimen on the microscope stage. Both the translationalmotion and intensity of the illumination source are remotelycontrollable under software program control operating as an applicationon the control processor.

A plurality of objective lenses 18 are connected to a rotatableobjective frame such that a specimen may be viewed at variousmagnifications. The rotatable frame may also be robotically controlledsuch that the various objective lenses can be moved into the microscopeoptical path and a specimen be viewed under any one of a number ofmagnifications at the desire of the operator. Examples of roboticallycontrolled microscopy systems suitable for use in connection with thepresent invention include the Olympus Vanox microscope system equippedwith a Prior H100 remotely controllable stage, or other similarcomputerized stages such as those manufactured and sold by Opelco.

A control processor, indicated at 20, implemented as a small platformcomputer system such as an IBM-type x86 personal computer system,provides the data processing and platform capabilities for hosting anapplication software program suitable for developing the necessarycommand and control signals for operating the microscope system. Thecontrol processor 20 is able to receive and interpret commands issued bya system user on a conventional input device, such as a mouse or akeyboard, and convert user defined commands into signals appropriate formanipulating the various components of the microscope system. Thecontrol processor 20 is typically coupled to the microscope systemthrough an interface, such as an SCSI interface, a proprietary interfaceor any one of a number of alternative coupling interfaces, which, inturn, defines a system bus to which the various control electronicsoperating the microscope system are connected.

A magnification control system suitably comprises a roboticallycontrollable motor and motor driver combination which is coupled to theobjective frame and is configured to rotate the frame to bring variousdesired objective lenses into the optical path. Upon receipt of anappropriate movement command signal, the magnification control systemdirects the motor to rotate the rotatable frame, thus moving a differentobjective lens into the optical path of the microscope system. Stagemovement is likewise robotically controlled by a stage movement controlsystem 16 which also comprises motors for moving the sample stage 14 inthe X, Y (16) and Z (17) directions along with appropriate motor drivercircuitry for actuating the motors. The mechanical apparatus andelectronic control circuitry for effecting stage movement is preferablyimplemented to include some form of open or closed-loop motorpositioning serving such that the sample stage can be either positionedwith great precision, or its translational movement can be determinedvery accurately in the X, Y and Z directions. For reasons that will bedescribed further below, it is important that the microscope stagerespond precisely to movement commands and that stage movement can bevery precisely determined and carefully controlled.

When the stage control system is configured to operate closed-loop,position feedback information can be recovered from the motor itself, orfrom optical position encoders or laser interferometer positionencoders, if enhanced precision is desired. Closed-loop servo control ofstage motion allows the stage position to be determined with greataccuracy and insures that translation commands are responded to withgreat precision. Thus, a command to translate the stage 50 microns inthe positive X direction will result in the stage moving precisely 50microns in +X, at least to the mechanical resolution limits of the motorsystem.

If the system is configured to operate semi-closed-loop, or open-loop,stage control is not dependent on feedback per se, but it is at leastnecessary to precisely define where the motors controlling the stagewere told to go. For reasons detailed further below, the transmittedvideo image is compression packaged in a manner that takes stage motion(both relative degree and absolute magnitude) into account when definingthe compression packaging technique used under various circumstances.

Focusing is performed by causing small excursions of the stage in the Zdirection under control of corresponding focus control circuitry 17.Because the amount of relative motion during focusing is significantlysmaller than the amount of relative motion during gross Z translationalmovements, the focus circuitry may well comprise a microstepping motorcontrolled by appropriate motor driver circuitry and operating inparallel with the Z axis stage translation motor. The z axis translationmotor could, thus, be provided with a more gross response characteristicso that it would be able to accommodate vertical optical sectioning of aspecimen, i.e., viewing a specimen on various horizontal planesvertically disposed therethrough, while the focusing motor wouldaccommodate the micromotions required to adequately focus each imageplane. Illumination circuitry controls the intensity of the illuminationlight source in conventional fashion.

A video camera 22 is optically coupled to the microscope to capturediagnostic-quality images of microscopic tissue samples disposed on thesample stage. The video camera 22 is preferably a high resolution,color, digital video camera operating at an image resolution inaccordance with, at least, the NTSE standardized composite color videoimage specification. Examples of video cameras suitable for use inconnection with the present invention include the Sony DKC-5000 seriesof video cameras, the Ikegami 370-M video camera, and other makes andmodels of composite, color video cameras of comparable quality andresolution. Images captured by the video camera 22 are directed throughvideo image processors 24 whereby they are able to be displayed on ahigh resolution digital display screen coupled to the telemicroscopecontrol processor 20.

In addition, and in accordance with the present invention,high-resolution video images captured by the camera are compressionpackaged by the control processor 20 for transmission over atelecommunications interface 26, coupled, in turn, to awide-area-network 28 such as the Internet.

The images developed by the video camera should have at least theresolution available under the NTSC standard. In North America andJapan, the NTSC color video image comprises approximately 480 pels perimage scan line in the red, green and blue (RGB) color components.Approximately 480 scan lines comprise an image frame, and image framesare generated at a rate of approximately 30 frames per second. If eachcolor component is coded as an 8-bit value (24 bits/pel, i.e., “truecolor”) representing some form of luminance plus color differenceencoding, for example, the representing continuous composite video isproduced at a bit rate of about 168 Megabits per second (Mbps). Fortruly high resolution image production suitable for telepathologyapplications, the video camera should be able to generate still imageswith resolutions preferably in the range of from 800×600 (pels×lines) toabout 1024×1024 (pels×lines). These images are transmitted at framerates of 30 fps and, at 24 bits per pel, this results in an image bitrate in excess 750 Mbps.

Clearly, the effective bandwidth of the electronic telecommunicationsnetwork connection which interconnects the host server system with apathologist's remote viewing and diagnostic facility (termed herein aclient system), determines the amount of time, or intrinsic delay,required to remotely display transmitted high resolution images on ahigh resolution monitor in a form and of a quality suitable fortelepathology applications. Ideally, the system and method of thepresent invention are implemented as an application software program,hosted on both a hospital's host server and a pathologist's clientsystem. The application program should be independent of thecommunication links interconnecting various sites, and should be capableof operation over a wide variety of conventional local and wide areanetwork (LAN and WAN) architectures. Preferably, the system isconfigured for optimum performance as a Windows-based TCP/IPimplementation, compliant with the WinSock 1.1 specification, and isthus compatible with LAN-over-broadband telecommunications architecturessuch as an Asynchronous Transfer Mode (ATM) based optical fiberconnection.

Basic rate ISDN, bundled ISDN, Asymmetric Digital Subscriber Loop (ADSL)and various Frame Relay telecommunications links are also contemplatedas electronic communications interconnections suitable for use inconnection with the system present invention. Although ATM-basedcommunication protocols in conjunction with an Internet-basedinterconnect architecture is preferred, all that is really required is acommunication architecture (connect hardware and data transmissionmethodology) able to bi-directionally communicate digital data atbitrate appropriate to the invention. The actual communication bandwidthrequired will, of course, depend on the kind and amount of compressionapplied to the raw video data stream. As will be developed in greaterdetail, below, compression packaging the video data stream in accordancewith the invention results in effective bit rates sufficiently lowenough to make effective use of many LAN/WAN architectures.

FIG. 2 depicts, in semi-schematic block diagram form, an exemplaryclient system which would typically be employed in the clinical practicefacility of a pathologist. The client system simply comprises a typicalpersonal computer system 30 equipped with a large, preferably 17 inch orlarger, high-resolution monitor system, 32, operatively controlled by ahigh-resolution, high-quality video graphics display subsystem hosted bythe system's electronics housing 34. A suitable implementation of theclient PC system would include a high speed processor, such as the Intelx86 series processor, sufficient random access memory to host thenecessary application software, a mass storage device such as a harddisk drive, and communications interface circuitry for effecting directelectronic communication with the server application of FIG. 1.Optionally, the client system might be provided with a videoconferencing suite including a video conferencing camera, a microphone,speakers and electronic interface circuitry, such that the client useris able to effect bi-directional audio/visual communication with theserver site in parallel with the sample video transmission.

Ideally, the pathologist's client system would operate within a Windows®or Windows-like graphical user interface (GUI) based environment inorder to best take advantage of the preferred Internet-basedbi-directional communication system in accordance with the invention.Operating within the familiar graphical Windows-based environment, thepathologist is able to control the remote telemicroscopy system bymerely “clicking” on various commands available through menus, orsubmenus, presented by the application software in a manner which mimicsconventional Webbrowser application software. Alternatively, the GUIenvironment would allow image tracking and movement commands to begenerated by “clicking” directly on the screen image. All of thetelemicroscope controls are available to the pathologist by using simplemouse commands, or alternatively, by issuing simple step commands over akeyboard. Multimedia functionality allows for bi-directional audiocommunication between the pathologist at the remote site and clinicalpersonnel at the telemicroscopy server, so that the pathologist is ableto converse with laboratory technicians, issue directions, makesuggestions and the like, all in parallel with the video feed. TheWindows® or Windows-like interface for microscope control and imagedisplay remains active throughout the entire session, for continuousease of use.

The methodology of the present invention is preferably implemented as anapplication software program running on both the telepathology networkserver and a pathologist's client system and is adapted to both controla telemicroscopy system and receive information from a video cameraconnected thereto. A microscope slide containing an investigationsection is placed on the robotically controlled sample stage of theserver system's telemicroscope. In one particular embodiment of theinvention, the server system might initially create a “mosaic” ofsmaller images which are arranged in a grid, or tiled, to define acomplete low power image of the entire tissue specimen to be examined.As a pathologist initiates an electronic communication with the serversystem, the mosaic image of the sample becomes available and is storedon a mass storage media device such as a hard disk drive on both systemsfor future reference. The pathologist may use the mosaic image as areference image or “roadmap” for further evaluation of the sample and isable to select any area of the mosaic for viewing at higher opticalmagnification. As will be described in greater detail below, the mosaicis initially transmitted with a particular level of detail, i.e.,magnification resolutions of from 1× to about 10× are immediatelyavailable for viewing. As additional resolution is desired, resolutionsgreater than 10×, for example, only the additional resolution detailrequired for examination of that particular mosaic “tile” istransmitted, representing a significant bandwidth savings.

As the pathologist determines which portion of the mosaic toinvestigate, higher optical magnification is obtained by issuing theappropriate commands over the client system, which are transmitted tothe server system through the network's communication connection.Additional portions of the mosaic are captured for viewing by issuingthe appropriate commands to move the remote sample stage of thetelemicroscope to appropriate X-Y positions which correspond to aselected location on the low power mosaic image. Selection of particularlocations for further viewing might be accommodated by “clicking” on theselected location using a mouse and further “clicking” on a desiredmagnification factor provided on a menu. All images transferred to theclient system remain on the display until no longer required. All imagesmay further be archived in a database along with their X-Y coordinateswith respect to the mosaic image, a time stamp, illumination data, andany such other information required to reconstruct the pathologist'sdiagnostic “trajectory” through the specimen. These archived images areparticular useful in developing a database for clinical educationpurposes.

Once the pathologist has determined where to begin examination of thespecimen, sample magnification is increased to an amount appropriate forexamination of detail and transmission of detailed video data isinitiated in accord with the invention. Video data is transmitted to theclient system according to an adaptive, multi-tiered compressionmethodology which dynamically adapts the form, content and detail oftransmitted images to the visual perception abilities of a typicalsystem user. To summarize, if a user is unable to perceive image detailbecause, for example, the image is moving due to sample stagetranslation, the image is adaptively transmitted at a significantlylower resolution, i.e., using significantly lower bandwidth. As theimage stabilizes, and higher resolution is perceptible, the systemadaptively increases transmitted detail. Thus, the system inherentlydelivers detail proportional to a user's ability to perceive that levelof detail.

The system is termed multi-tiered, in that various forms of compression,both cognitive and operative, are used to define the transmitted imagedata. For purposes of illustration only, an exemplary embodiment of theinvention will be described in terms of a three-tiered compressionsystem in which the tiers comprise; first, necessary image transmission;second, progressive image encoding, and; third, bitwise imagecompression, exemplified by the JPEG still image compression standard.

In operation, when a pathologist selects a particular sample portion fordetailed viewing and requests the sample stage to move in the X or Ydirection, the server system controls stage movement according toappropriate commands received from the client via a special-purposeTCP/IP network protocol. It should be noted, here, that this networkprotocol is merely an extension of HTTP and is configured to supportapplication-specific protocols relating to session management,hierarchical image decomposition, image compression, data transmissionand telemicroscopy control commands. These commands include translatingthe sample stage in the X or Y direction in order to alter the relativeX-Y position of the camera with respect to the stage and, thus, thespecimen or frozen section. When the pathologist requests stage movementin the X or Y direction, the client system issues this request to thetelemicroscope through the server, the telemicroscope system moves thestage and a new portion of the section is brought into view. It isimportant to realize that when the sample stage is moved, the priorimage is effectively translated in the X and/or Y directions by aspecific amount, defined by the pathologist working at the clientsystem.

As illustrated in FIG. 3, as an image is translated, the resulting newimage may be decomposed or defined in terms of two image portions; afirst portion 36 comprises those parts of the prior image which remainin the field of view but have been translated by an amount determined bythe X and Y stage motion commands (termed herein previously transmittedportions) and those portions of the image 38 which have been translatedinto the field of view and which constitute previously unseen and, thus,untransmitted information (termed herein new portions). In accordancewith the present invention, the client system is able to calculate towhat degree the previously transmitted image portions must be displacedin the X or Y direction in order to reflect the translational movementcommands directed to the sample stage. Those previously transmittedportions are merely displaced in the display system with those sectionswhich presently fall outside the field of view being “dropped-off” theedges in the direction of translational movement, as indicated in FIG. 3at 40.

It will be understood by those having skill in the art that thosepreviously transmitted image portions 36 which remain in the presentfield of view need not be retransmitted by the server system, but needonly be analyzed and displaced by the client. Those portions of the newimage which were not previously transmitted, i.e., the new portions 38,are the only image portions which require transmission. This necessaryimage transmission procedure results in a significant bandwidth savingswhen translational motions in both the X and Y direction are relativelysmall, requiring a significantly reduced amount of image informationtransmission in order to define the new field of view. Once the newimage portions have been determined, those image portions are designatedfor transmission and may be either directly transmitted in a PKZIP-typebitwise compression format, for example, or alternatively, compressionpackaged in accordance with the system and method of present inventionand subsequently transmitted as a compressed bitstream.

The progressive encoding tier of the multi-tiered compression scheme istypically invoked when sample stage translational movement is fastenough that entire frames, or substantial portions of frames are“dropped off” such that the majority of the image comprises a “new”image portion. In this case, the image is subject to a progressiveencoding scheme based on differential subsampling. In accordance withthe invention, image sampling would only be performed with respect toevery other pel, for example, in both X and Y. Taking only half the pelsin both the X and Y dimensions, typically results in a reduction in theimage size by about 75%. However, overall image size can be maintainedby merely replicating each pel in software, at the receiving (client)site, resulting in a full-size image having one-fourth the originalresolution density. In a 1024×1024 system, this factor-two progressiveencoding scheme would result in an image having the effective resolutionof a 512×512 original. This resolution level is certainly acceptable forfast panning and is also marginally acceptable for “quick-stop” fastimpressions.

Once panning is concluded (i.e., when the system no longer “sees” Xand/or Y translation commands being issued to the server), the unsentportions of the video signal are transmitted in background, and are usedto fill-in the image in place of the replicated pels, until the ideal1024×1024 resolution is reached. Needless to say, the subsampling indexcan be selected by the user through a settable system parameter. Theprogressive encoding tier is able to be programmed to sample every otherpel, every third, every fourth, etc. according to the perceptual desiresof the user. Lower value indices, while increasing resolution duringfast panning, do so at the cost of bandwidth and should be invoked onlyafter careful consideration.

Turning now to FIG. 4, which is a semi-schematic block diagramrepresentation of a bitwise compression encoding and packaging engine inaccordance with the practice of principles of the present invention,video data representing a frame of video information is captured by avideo camera and initially provided to a frame segmentation anddifferencing engine 42. Briefly, the frame segmentation and differencingengine 42 is responsible for the first compression step, i.e.,identification of those portions of the video frame which are “new” and,thus, necessary for transmission. Frame segmentation and differencingcan be best understood as the process of comparing a present video framewith an immediately previous video frame, represented at 44, anddetermining which portions of the previous frame are to be retained,which portions of the previous frame are to be discarded fortransmission purposes and which portions of the present frame representnew image data requiring transmission. With regard to identifying thoseportions of the previous frame which are to be retained, the framesegmentation and differencing engine 42 is further able to define thedisplacement vector difference between the present and previous framesby analyzing position and focus change commands directed to thetelemicroscope system.

In accordance with the invention, the previous frame video data 44 isprovided to the frame segmentation and differencing engine by retrievingthe previous frame data from server system memory and decompressing thedata using a decompression engine 46 to, thereby, redefine the raw videoinformation defined in the previous frame. Previous frame data is“overlaid” to the present frame data in software to define those “new”portions which require transmission.

Following frame segmentation and differencing, the “new” portions of thepresent video frame data is compressed, in a transform block 48, inaccordance with any one of a number of lossy compression techniques,such as the Discrete Cosine Transform (DCT), as implemented in the JPEGcompression standard, wavelet transform coding, and the like. Whetherlossy compression is performed using Discrete Cosine Transforms orwavelet transform coding, the techniques result in the video image beingrepresented as a set of coefficients representing the contributions ofvarious frequency components of the signal. Once decomposed into theirvarious frequency components, frequency component coefficients are“packetized” and inserted into a number of transmission queues 50 which,in a manner to be described in greater detail, are assigned totransmission priority levels based on the level of detail which eachpacket additively contributes to the resultant composite image. In thecase of the Discrete Cosine Transform or a wavelet transform, the lowerfrequency components represent the least amount of image detail andpackets which define lower frequency component coefficients are,accordingly, assigned the highest transmission priority. As detail, andthus, frequency, increases, packets containing higher frequencycomponent coefficients are assigned correspondingly lower transmissionpriorities until, in the limit, the highest levels of detail (thehighest frequency components) are assigned the lowest transmissionpriority.

Packetization and prioritization of transform encoded frequency data isan important aspect of the present invention, since it allowstransmission bandwidth to be adaptively varied in order to accommodatethe subjective, perceptual aspects of a moving image system. Forexample, when sample stage motion is being executed quickly, i.e.,relatively large amounts of X and/or Y translational movements, onlythose coefficient packets representing the lowest frequency componentsof the image, are transmitted to the client system for display. It willbe understood that if stage movement is rapid, a viewer is unable toperceive fine detail making it unnecessary to burn bandwidth or channelcapacity attempting to transmit volume data relating to imperceptibledetail. It is understandably more important to service the client systemwith faster overall frame updates which might be detail deficient thanit is to service the client with specimen detail at considerably slowerintervals.

As stage movement is slowed down, i.e., as X and/or Y translationalmovements become less, additional transmission queues are flagged fortransmission and bandwidth that is not being used to send new fields ofview is available for transmission of additional packets comprisinghigher frequency component coefficients for the current view. Adding theinformation from additional transmission queues necessarily enhances thedetail of the current image being displayed by the client system.Necessarily, as translational movement of the sample stage ceases, allthat remains is for the final transmission queues, comprising thehighest detail level component coefficients, to be flagged fortransmission to the client system.

The information contained in these final transmission queues may makeuse of all of the available transmission bandwidth since lower frequencyinformation (higher priority packets) have been previously transmitted.It will be understood by those having skill in the art that image detailis accordingly inversely proportional to stage translational movementspeed, with higher speed movement resulting in lower image resolution.Thus, the compression encoding procedure in accordance with theinvention is able to inherently deliver image resolution or detail inproportion to the resolution level that is able to be perceived by aviewer. Compression encoding and adaptive prioritization of imageresolution packets defines an image system that is able to efficientlybuild upon previously transmitted data in order to minimize transmissiontime (maximize bandwidth) for a full resolution image.

Returning now to FIG. 4, transmission queues 50 containing prioritizedfrequency component packets, are optionally preceded by components thatare configured to provide an additional degree of either lossy, orlossless compression encoding to each of the respective componentcoefficient packets defined by the transform engine. These optionalcomponents might suitably comprise quantization engines 52 followed byarithmetic coding or entropy coding engines 54. Both quantization andarithmetic or entropy coding are options available in the JPEG standardand need not be described in detail herein. Specifically, baseline JPEGuses Discrete Cosign Transform compression of 8×8 blocks of pels,followed by scalar or vector quantization of the DCT coefficients andentropy encoding of the result. JPEG specifies two possible entropyencoders, one of which is based on the well understood Huffman codingalgorithm and which is required for the baseline JPEG system. Extensionsto the JPEG standard and JPEG lossless compression modes also permit theuse of arithmetic coding, exemplified by the IBM Q-Coder, as analternative.

Compressed video image data is then moved through the server systemwhence it is transmitted to the client application over a broad bandtelecommunications architecture via an ATM-based communication channel56 using, for example, IP-over ATM LAN emulation. While transmissionqueues are forwarding various components of the video image data to theclient system, the server application is able to receive telemicroscopestage translational motion commands and/or focus commands from apathologist at the client, through translation command circuitry 58 andfocus command circuitry 60. If a particular stage translational movementis requested that moves a portion of the current image out of the fieldof view, coefficient packets that correspond to those blocks comprisingthat portion of the image are flagged with an identification tag whichindicates that these packets are no longer in the field of view and,accordingly, need not be transmitted.

It will be understood, of course, that in the case of Discrete CosineTransform compression, block sizes will be in the familiar 8×8 form. Inthe case of wavelet transform compression, wavelet blocks are variablysized, and typically larger than the 8×8 DCT standard. It will beunderstood by those knowledgeable in the art, that common practice wouldnormally be to apply the wavelet across the entire new image portionstrip, thereby defining the block size as the new image portion. Whilethis practice maximizes compression efficiency, it does so at theexpense of packetization and prioritization efficiency. It should befurther understood that the new image portion can be subdivided intosubimages with each of the subimages wavelet transformed in order toimprove efficiency of the flagging process (packetization andprioritization). Necessarily, this will be at the expense of somecompression efficiency. Ideally, wavelet blocks are sized variably inaccordance with a design choice that achieves a desired balance betweenout-of-view flagging and wavelet transform performance efficiency.

In the case of changes to specimen focus, i.e., stage motion in the Zdirection, large portions of the currently displayed image undergosubstantial modification. The new image, resulting from a focus changeor a change illumination intensity, is subtracted from the previouslytransmitted image represented by the previous frame data in the framesegmentation and differencing engine, resulting in an image comprised ofthe difference between the previous frame and present frame as seen bythe video camera. The difference image is then transform coded by thetransform engine and the resulting lossy compression packaged data isthen optionally quantized and/or entropy or arithmetically coded fortransmission.

As is shown in the semi-schematic block diagram of FIG. 5, transmittedvideo data is decompressed in accordance with an inverse sequence ofprocesses and the resultant inverse transform data is summed to definethe current video frame. In a manner similar to frame segmentation anddifferencing, a composite picture 66 is built up by adding current framedata 62 to retained portions of the previous frame data 64 which havebeen displaced, in software, in accordance with any translational motionor focus vector changes. The resulting composite frame 66 is then sentto the videographics client for rendering on the display screen.Received compressed video information is decoded in respective inversearithmetic coding engines, from whence the data is directed to inversequantization engines 70. Each packet is then reconstructed in accordancewith an inverse transform, in respective inverse transform engines 72,and summed, in a summing circuit 74, to become the current framerepresentation 62 of the image.

It has been determined that the multi-tiered compression packaged imagetransmission system in accordance with the invention is able to delivervideo image data at an effective compression rate of approximately1:400. Given a raw video input signal comprising a 1024×1024 pel×frameproduced at a frame rate of approximately 30 frames per second, witheach pel represented by a 24 bit (true color) digital value, the rawdata stream would be operative at approximately 755 Mbps. Assuming a ⅔screen width pan taking place in 0.25 seconds, the multi-tieredcompression system according to the invention would be providing videodata to the client system at an effective rate of about 1.65 Mbps whenall of its component portions are considered.

The necessary image transmission portion of the novel compression systemaccounts for at least an order of magnitude reduction in the effectivebit rate; 755 Mbps reduced to 66 Mbps. Transform coding andprioritization queuing of the resultant output stream results a further40 to 1 reduction in the effective bit rate; from 66 Mbps to about 1.65Mbps, with progressive encoding accounting for a 4:1 reduction in thebitstream (from 66 Mbps to about 16.5 Mbps). It should be noted thatthis last value represents a two-fold improvement over the generallyaccepted standard JPEG capabilities. Without being bound by theory, itis thought that the major portion of the improvement results from theability of the novel system to adaptively deliver image detail ininverse proportionality to sample stage movement speed and in directproportionality to the degree of detail that can be perceived, while atthe same time, building detail upon previously transmitted data fortransmission bandwidth utilization efficiency.

The necessary image transmission portion of the novel compression systemaccounts for at least an order of magnitude reduction in the effectivebit rate; 755 Mbps reduced to 66 Mbps. Transform coding andprioritization queuing of the resultant output stream results a further40 to 1 reduction in the effective bit rate; from 66 Mbps to about 1.65Mbps, with progressive encoding accounting for a 4:1 reduction in thebitstream (from 66 Mbps to about 16.5 Mbps). It should be noted thatthis last value represents a two-fold improvement over the generallyaccepted standard JPEG capabilities. Without being bound by theory, itis thought that the major portion of the improvement results from theability of the novel system to adaptively deliver image detail ininverse proportionality to sample stage movement speed and in directproportionality to the degree of detail that can be perceived, while atthe same time, building detail upon previously transmitted data fortransmission bandwidth utilization efficiency.

While the foregoing has included a description of the present inventionin connection with an exemplary telemicroscopy application, it should beunderstood that principles of the invention are equally applicable toimage transmission methodologies that encompass various combinations oflossy and lossless image compression, particularly when adapted forimage transmission and archiving purposes. Taking the principles of theinvention into account, i.e., image transmission and display ispredicated upon the ability of a viewer to perceive the degree of detailpresent in a particular image, image compression is subdivided intolossy and lossless image compression stages, with image portions beingtransmitted from various queues in accordance with viewerperceptualability. In the present context, lossless image compression iscommonly understood to refer to compression techniques that allow aparticular image, when decompressed, to be replicated precisely, at abit-by-bit comparison level, in the same form as the original image.Particular types of lossless image compression algorithms might includerun length encoding (RLE), Huffman coding, dynamic Huffman coding,Lempell-Ziv-Welch encoding, and the like. While many losslesscompression techniques do afford some degree of compression, i.e., somedegree of bandwidth recovery, lossless compression techniques do nottypically afford sufficient bandwidth recovery for rapid imagetransmission over commonly implemented transmission channels. Thus,lossless image compression, while accurate, is slow.

Lossy compression techniques result in a recovered image that is notprecisely the same, at a bit-by-bit comparison level, as the originalimage. Lossy compression techniques result in a significantly higherdegree of bandwidth recovery than lossless techniques, but the amount ofprecision detail available in the resultant image is often insufficientfor high magnification applications such as telemicroscopy, and thelike.

With the advent of Internet-type wide area networking, many applicationsthat were previously limited to local use, i.e., direct high-speedaccess to a local memory storage area such as a local hard drive, arenow being implemented by systems separated by great distancesgeographically. Many of these applications, such as fluorescent imaging,CAT imaging, and the like, involve quantitative analysis of the image.Accordingly, it is necessary that the final image remain “lossless” uponcompression without unduly interfering with bandwidth constraints ofremote viewing and data archiving.

Turning now to FIG. 6, a particular advantage of the principles of thepresent invention is that lossless compression of images for botharchiving and transmission purposes may be achieved without sacrificingtransmission speed when images are accessed by a remote location. As canbe seen from the exemplary embodiment of FIG. 6, lossy and losslesscompression techniques are layered, with the various layers representingincreasing amounts of detailed structure available for a particularimage. In FIG. 6, the coding layers are divided into a lossy layerportion 100 and a lossless layer portion 102. The lossy layer portion100 is further subdivided into a plurality of layers, represented byordinal identifiers ranging from 1 to N−1, as indicated in FIG. 6.According to the invention, lower ordinal layers 1 to N−1 are adapted tocontain data obtained from a lossy compression methodology such as adirect cosine transform (DCT), a wavelet transform, fractal compression,and the like, as was described above. Coefficients arising from suchtransforms are quantized and prioritized into the various lower ordinallossy layers 100, as was described previously in connection with theembodiment of FIG. 4. Prioritization, necessarily, is expected to varyfrom application to application, with coefficients being typicallyclassified in accordance with theirs importance in assembling the finalimage. In the specific case of wavelet coding, lower order coefficients,representing low frequency data, are prioritized into lower-valuedordinal layers, such as layer 1, layer 2, or the like, while higherfrequency information (higher order coefficients) representing detailinformation, are prioritized into higher and higher layers,corresponding to greater and greater amounts of detail.

As was the case with the embodiment of FIG. 4, following transformationand quantization, the various layers are arithmetically coded andentered into an appropriate transmission queue 104 for transmission to aremote location over WAN transmission channels.

In order to retain fidelity with the original image, a lossless portion102 is provided for containing image data that, when combined with lossylayer information, results in a lossless image. The lossless portion 102contains data difference information which is constructed by subtractingthe original image representation from the assembled image informationcomprising lossy layers 1 to N−1. This difference data is compressed bya lossless compression technique such as Huffman coding, dynamic Huffmancoding, or Lempel-Ziv-Welch coding in order to minimize its particulartransmission bandwidth requirements.

As was the case with the lossy layer portion 100, the lossless portion102 may also be implemented as a plurality of layers, each layerrepresenting perhaps a frequency code, in the case of Huffman coding, orsome other statistical weighting in the case of the various othersubstitutional or statistical compression coding methodologies.Similarly, the lossless information is directed to a respective transmitqueue 106, which is flagged as the final queue, i.e., transmitted afterall the lossy queues 104 have been streamed out. As was also the case inthe embodiment of FIG. 4, after receipt and decoding, the queue contentsare summed, in a summing circuit 108 to build a composite image 110.

As depicted in FIG. 7, the principles of the present invention offerseveral advantages over lossless or lossy transmission techniques alone.In particular, image sizes processed in accordance with the inventionwill be expected to be substantially the same as a losslessly compressedimage in accordance with LZW or Huffman statistical coding, whileretaining the rapid download and viewing capability of a lossycompression technique such as JPEG. The present invention realizes theseadvantages by the particular nature of its transmission queuing whichprovides the ability to stream an image to remote locations at a highdata rate with an attendant reduction in image detail for rapid viewing,while filling-in image detail with time. Thus, the present invention isable to deliver an image at a level of detail proportional to a visualperception level of a user. In the case of a large image, such as apathology slide, as a user scans the specimen, image information isaccessed from the higher priority transmission queues, i.e., thosequeues containing only lower frequency (lower detail) information.Transmission queue sequencing can be understood as being related tostage motion or frame translation, with transmission queuing recyclingto higher priorities as the image translates. “Fast panning” across animage only allows higher priority queues to be transmitted anddownloaded. As image translation speed is reduced, more and moretransmission queues are accessed thereby adding detail to the displayedimage. When an area of interest is identified, and frame translationceases, all layers queued for transmission are downloaded giving aresulting image substantially the same as the original opticallyobtained image.

It should be understood from the time graph of FIG. 7, that the lowerordinal layers (higher priority layers) have already been downloaded asa function of frame translation. Thus, as an image ceases to move, onlythe higher ordinal layers (lower priority queues) and the losslesslayers need be transmitted in order to reconstruct the final image.Thus, a clinician may rapidly traverse an image, looking at only thelower detail levels, and perceiving a near visually lossless image. Astranslation slows down, greater and greater amounts of detail areallowed to fill in as additional transmission queues, containingadditional detail layers, are accessed. Image detail, then, becomes acumulative time function, with transmission bandwidth requirementsremaining substantially level over time. The final image is thus theresult of a time integration, with the full-size image being constructed“vertically” by “overlaying”, or enriching, detail information onto alow resolution primitive, as opposed to the entire image's beingcaptured, compressed, transmitted and “painted” on a display.

When applied to an application such as pathology slide evaluation,principles of the present invention might be used in combination with animage “tiling” or mosaic creation technique such as describedpreviously. In this particular exemplary embodiment, very large imagesare segmented into a mosaic of tiles, i.e., a 16×16 pattern, forexample, each of which are processed in accordance with the transform,quantization, coding and queuing methodology of the invention.Specifically, compression might be lossless, lossy or the particularcombination of the two described above in order to minimize bandwidthutilization and improve download and viewing speed. The lossless/lossytransmission methodology described above, is particularly useful in thisparticular application. When a large, overall image is being initiallyconstructed, it is only necessary to create the mosaic tiles with anintermediate degree of detail, since a user's perceptual ability cannotdiscern fine structure at such low magnifications. Thus, each tile'simage information need only be processed by the lossy techniquesdescribed above and only the higher priority transmission queues (lowerfrequency data) need be transmitted. An image mosiac constructed in suchfashion might then support image detail sufficient to represent 2×, 5×and even 10× magnifications, such that rapid identification of areas ofinterest in the slide might be made at lower magnification levels, whilefine structure for those areas of interest remain untransmitted untilsuch time as a user requires the additional image information.

In this particular instance, image detail delivery is again proportionalto a user's perception level, but rather than being a function of frametranslation speed, it is a function of magnification level. At lowmagnifications (the entire image for example) sufficient detail for userperception is provided by the lossy compressed data contained in thehigher priority transmission queues. There is no need to provide finestructure in such circumstances, because the user is unable to provideit. Bandwidth is thereby conserved. As magnification increases, imagesize necessarily decreases. Accordingly, fine structure detail need onlybe provided for a small portion of an image at any particular time.Thus, fine structure detail is not required for the entire image, againconserving transmission bandwidth while not degrading the image.

A further advantage of the present invention can be understood when itis realized that it need not be limited to transmitting real-timevideographic images captured over a telemicroscopy system. Specifically,the system and method of the present invention has equal utility intransmitting and viewing “virtual specimen” videographic informationwhich has been previously captured by a computer controlledtelemicroscope and which has been stored in a server database as asequence of high-resolution images. When the digital nature ofinformation captured by a telemicroscope's video camera is considered,it will be understood that this information can be used to define amulti-dimensional mosaic of a sample specimen which may be “navigated”in precisely the same fashion as an actual sample. Virtual specimens canbe evaluated in detail by traversing the video file's virtual dimensionsin a manner quite similar to issuing X, Y and Z motion commands to atelemicroscope. When it is understood that the systems described areeasily capable of incorporating real-time video conferencingapplications suitable for connection to the described telecommunicationsinterfaces, one having skill in the art will immediately appreciate theutility of this system as a means for archiving rare or unusual tissuesamples, either for further review or future comparison purposes,consultations and educational presentations.

Although the present invention has been described in connection with aparticular illustrated embodiment, it should be understood that variouschanges, substitutions and alterations can be made without departingfrom the spirit and scope of the present invention. In particular, thenovel system has been described in connection with a telemicroscopyapparatus that incorporates a conventional optical microscope. It shouldbe realized that future instruments adapted to image microscopicspecimens may not be configured as microscopes in the traditional sensebut function rather more like desktop scanners having compound opticalsystems. Digital signals representing a microscopic image might notconform to the NTSC standard nor even be video signals as that term iscurrently contemplated. It should be understood by those having skill inthe art that the form and arrangement of digital signals representing amicroscopic image is not the primary concern of the invention, butrather represents an information package which is operated on by a novelcompression system and method.

There has now been brought to the field of telemicroscopy an improvedsystem and method for compression packaging video image data in a mannerwhich overcomes channel capacity limitations and allows almost real-timeevaluation of tissue samples. A flexible, variable and self-adaptingmethodology is used to compress and reduce the image datastreamaccording to the observation activity of a user.

1. A method of providing remote navigation of a specimen, comprising:receiving a command to remotely control a microscope and a cameracoupled with the microscope to view a portion of a specimen disposed onthe microscope; capturing a digital image of the portion of the specimenwith the camera in response to the command; using a processor torepresent the digital image as a plurality of digital image components,each of the digital image components providing a different level ofdetail of the image; and transmitting the components in sequential orderof increasing level of detail from least detailed to most detailed. 2.The method of claim 1, wherein the microscope comprises a stage, andwherein the specimen is disposed on the stage.
 3. A system providingremote navigation of a specimen, comprising a processor that executesinstructions and thereby causes the processor to: receive a command toremotely control a microscope and a camera coupled with the microscopeto view a portion of a specimen disposed on the microscope; capture adigital image of the portion of the specimen with the camera in responseto the command; represent the image as a plurality of digital imagecomponents, each of the digital image components providing a differentlevel of detail of the image; and transmit the components in sequentialorder of increasing level of detail from least detailed to mostdetailed.
 4. The system of claim 3, wherein the microscope comprises astage, and wherein the specimen is disposed on the stage.